摘要
目的探讨柯萨奇病毒(CVB3)心肌炎细胞毒T细胞(CTL)亚类(ACTL、VCTL、MCTL)TCR CDR3VΒ基因取用格局。方法无菌摘取72H内新生BALB/C鼠心脏,培养单层心肌细胞并将其分为三组,分别用CVB3、放线菌素D处理和不经过任何处理;实验组制备CVB3心肌炎鼠模型,处死后将其肠系膜淋巴结制成单细胞悬液,分别加入上述三组单层心肌细胞,采用免疫吸附法获取ACTL、VCTL、MCTL,对照组鼠为正常小鼠处死后取肠系膜淋巴结制成单细胞悬液;采用MTT法对实验组ACTL、VCTL、MCTL和对照组T细胞进行细胞毒鉴定;常规进行RT-PCR,判断实验组细胞及对照组细胞的TCR VΒ基因取用格局。结果对照组T细胞表达VΒ基因全部20个家族,而实验组三类CTL的VΒ基因取用格局呈现明显的限制性,ACTL优势表达VΒ6、VΒ8.1、VΒ8.2、VΒ8.3,MCTL优势表达VΒ5.1、VΒ8.1、VΒ8.2、VΒ8.3,而VCTL优势表达VΒ7、VΒ8.1、VΒ8.2、VΒ8.3。结论CVB3心肌炎中导致心肌细胞损伤的CTL受到特异性抗原刺激后其TCR VΒ基因取用格局呈现明显的限制性。
Objective To analyze the T cell receptor （TCR） Vβ gene usage of the cytolytic T-lymphocytes which result in immune-mediated myocyte injury in Coxsackievirus B3-induced myocarditis. Methods An experimental murine model with a myocarditic variant of Coxsackievirus B, type 3 has been developed with newborn Balb/c male mice. Mice were killed on day 7 after virus inoculated, which was the time when T cells show significant differentiation. With immunoadsorption, ACTLs, VCTLs, MCTLs are respectively isolated from mesenteric lymph nodes of Balb/c mice suffered from viral myocarditis. T-lymphocytes prepared from mesenteric lymph nodes of uninfected Balb/c mice served as control group. RT-PCR was performed with standard method. The Vβ gene usage was analyzed by running the electophoresis of PCR products on agarose gel. Results T cells of control group expressed all Vβ genes of 20 families. But the TCR Vβ gene usage of CTLs in experimental groups was markedly restricted. ACTL expressed predominantly Vβ6, Vβ8. 1, Vβ8. 2, Vβ8. 3, MCTL expressed predominantly Vβ5. 1, Vβ8. 1, Vβ8. 2, Vβ8. 3 and VCTL expressed predominantly Vβ7, Vβ8. 1, Vβ8. 2, Vβ8. 3. Conclusions The TCR Vβ gene usage of CTLs which result in immune-mediated myocyte injury in Coxsackievirus B3-induced myocarditis is markedly restricted in cells stimulated by specific antigen.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2006年第1期80-83,共4页
Journal of Sichuan University(Medical Sciences)
